1. Field of the Invention
The present invention relates to a novel xyloglucan oligosaccharide composition designated anti-microbial adhesion inhibitory fraction A6, a composition containing said oligosaccharide composition containing anti-microbial adhesion inhibitory fraction A6, and methods of using said composition to at least reduce the adhesion of pathogens to animal cells, especially human and mammalian cells.
2. Description of the Related Art
Xyloglucan is well known as a major cross-linking polysaccharide in type 1 plant cell walls found in dicotyledonous and non-commelinoid monocotyledonous plants (Carpita and Gibeaut, Plant Journal, Volume 3, 1-30, 1993). With a β-(1-4)-glucan backbone, xyloglucan hydrogen bonds to the surface of cellulose microfibrils and forms a network that connects adjacent microfibrils in cell walls. The xyloglucan network is intermeshed with the pectin network of cell wall matrix polysaccharides (Carpita and Gibeaut, 1993 supra). This makes xyloglucan an important polysaccharide in the growth and development of primary cell walls (Carpita and McCann, “Biochemistry and Molecular Biology of Plants, Buchanan B. B., Gruissem, W., Jones, R. L., Eds.; American Society of Plant Physiologists, Rockville, Md., 52-108, 2000). There is a block-like structure in xyloglucan where a 6-11 sugar sequence is repeated throughout the polysaccharide. Therefore, carbohydrate structures are specific for plant taxonomic groups (Sims et al., Carbohydrate Research, Volume 293, 147-172, 1996; Vierhuis et al., Carbohydrate Research, Volume 332, 285-297, 2001; Ray et al., Carbohydrate Research, Volume 339, 201-208, 2004; Hoffman et al., Carbohydrate Research, Volume 340, 1826-1840, 2005). Three types of xyloglucan structures have been described with fucogalacto-xyloglucan the most commonly distributed in about half of the monocot taxonomic orders and all dicot orders except for the Solanales, Laminales, Gentianales and Ericales (Carpita and McCann 2000, supra; Hoffman et al., 2005, supra). Xyloglucan from these later orders contains arabino-xyloglucan structure. Small amounts of a third xyloglucan structure are also present in commelinoid monocots (grasses, bromeliads, palms, and cypresses) as randomly distributed single xylose substituents on a cellulosic backbone (Carpita and McCann, 2000, supra). A single letter nomenclature was developed to describe the sequence of xyloglucan substituents (Fry et al., Physiol. Plant., Volume 89, 1-3, 1993).
Cranberry juice is acidic (pH approximately 2.6 or lower) and rich in anthocyanins and tannins giving it an astringent taste (Holmes and Starr, Fruit Juice Processing Technology, Nagy, S., Chen, C. S., Shaw, P. E. (Eds.), AGSCIENCE, Auburndale, Fla., 515-531, 1993). The juice is prepared by milling and pressing after a hot (approximately 50 degree C. for about 1 hour) commercial pectinase maceration of the berries. Cranberry pectin has very high methoxy content, which requires a second hot commercial pectinase treatment following pressing and prior to juice filtration and concentration. Cranberry juice is considered a healthy juice. The proanthocyanidins have antioxidant properties (Uri-Sarda et al., Anal. Bioanal. Chem., Volume 394, 1545-1556, 2009) and were reported to inhibit adhesion of p-fimbriated Escherichia coli to uroepithelial cells (Howell et al., Phytochem., Volume 66, 2281-2291, 2005). P-fimbriated E. coli is the major cause of urinary tract infections which result in 8.3 million doctor office visits per year (Zopf and Roth, Lancet, Volume 347, 1017-1021, 1996). Cranberry juice was also reported to have prebiotic properties (Clifford et al., U.S. Patent Application No. 20090022849, 2009).
Recently, Coleman et al. (Presentation on Jul. 13, 2010 to the American Society of Pharmaconosy, St. Petersburg Beach, Fla.) fed pigs cranberry juice powder and isolated oligosaccharides or aminosugars from the urine that inhibited red blood cell agglutination using uropathogenic E. coli. The same α-Gal-(1-4)-β-Gal receptor is required for red blood cell agglutination and p-fimbriated E. coli adhesion to uroepithelial cells (Howell et al. 2005, supra). Therefore, a carbohydrate, not derived from proanthocyanidins, was reported to have bacterial anti-adhesive properties (Coleman et al. 2010, supra). Pectic oligosaccharides inhibited the adhesion of verotoxigenic and enteropathogenic strains of E. coli to HT29 cells (Rhoades et al. J. Food Protect., Volume 71, 2272-2277, 2008). So there is precedence for plant cell wall oligosaccharides to have bacterial anti-adhesive properties. However, the structure for cranberry plant cell wall oligosaccharides is unknown and Coleman et al. (2010, supra) did not report how an oligosaccharide was absorbed in the gastrointestinal tract prior to urinary excretion.
Current therapeutic regimes for the neutralization and/or removal of bacteria and bacterial components from host organisms, such as humans and domestic livestock, are based largely on the use of antibiotics. Since their introduction in the 1940's, antibiotic drugs have proven effective for the treatment of many bacteria-related illnesses. However, their frequent misuse has given rise to antibiotic-resistant bacterial strains that have necessitated the development and implementation of increasingly more powerful drugs. Bacterial infections are increasing within hospitals, resulting in a dramatic increase in cases of patients with serious, and in some instances, life-threatening symptoms. By way of example, urinary tract infections (UTIs) have been a pervasive health care problem. UTIs are generally defined as the presence of >100,000 cells/mL of bacteria in the urine. UTIs are commonly caused by Gram-negative bacteria, particularly Escherichia coli (E. coli), and infect primarily women. This infection is enabled by the adherence and colonization of bacteria to urinary tract epithelial cells. Adherence by E. coli is performed by proteinaceous fibers (fimbriae) on the bacteria cell wall, which attach to specific oligosaccharide receptors on uroepithelial cells. Antibiotics are commonly prescribed for treatment, but often promote bacterial resistance. One in four women also encounter recurrence of the infection and are often found to be prone to such infections. Natural substances which could treat and prevent UTIs could be useful for those suffering this condition since antibiotic treatment, in many cases causes, as secondary vaginal yeast infection requiring a subsequent antifungal treatment.
There is a large literature on the role of cranberry phytonutrients in preventing or mitigating urinary tract infections (UTIs), and particularly the Gram negative uropathogenic bacterium E. coli, the most common cause of UTIs (Lavigne et al., Clinical Microbiology and Infection, Volume 14, 350-355, 2008; Ofeck et al., Advances in Experimental Medicine and Biology, Volume 408, 179-183, 1996; Ofeck et al, New England Journal of Medicine, Volume 324, 1599, 1991). Consumption of cranberries has been found to be somewhat effective in addressing UTI infections. Cranberry products can prevent adhesion of certain bacteria fimbriae to uroepithelial cells in the urinary tract, thereby reducing the ability of the bacteria to create an infection (DiMartino et al., World Journal of Urology, 2006); (Liu et al., Biotechnology Bioengineering, 2006). Proanthocyanidins, which are condensed tannins, found in the cranberry juice have been shown to inhibit E. coli adherence (Howell et al., Journal of Medicine, 1998). United States Patent Application 2009/0226548, published in 2009, states that some E. coli fimbriae bind specifically to D-mannose, unlike sucrose or fructose, which is metabolized very slowly in humans, therefore once consumed, D-mannose will enter the blood stream and quickly moves to excretion via the kidneys followed by entry into the bladder in urine. D-mannose once in urine will cause the bacterial fimbriae sensitive to D-mannose binding to attach to the D-mannose, rather than epithelial cells. This allows the body to flush the D-mannose bound E. coli bacteria from the body. In addition, D-mannose can reverse epithelial bound E. coli competitively interrupting the initial phases of urinary tract infection. To mitigate existing UTIs and prevent recurrence, regular consumption of cranberry in combinations with D-mannose will prevent bacteria from adherence, colonization and ultimately prevent an uncontrollable urinary tract infection. This reference further states that for this strategy to work, consumer compliance is necessary,
Another treatment for bacterial and fungal infections is the use of cinnamon (Cinnamonmum cassia) extracts. The antimicrobial action of cinnamon can be partly attributed to the presence of cinnamaldehyde, eugenol, borneol, linool, and thymol, mainly antibacterial, and o-methylcinnamaldehyde, mainly antifungal.
There remains a need for nutrition and therapeutic compositions and methods for at least reducing inhibition of bacterial adhesion to human and mammalian cells and for reducing or inhibiting the invasion and infection of human and mammalian cells by pathogenic bacteria. The present invention described below includes such compositions and methods which are different from related art compositions and methods.